Abstract
Besides astrocytes and oligodendrocytes, NG2 proteoglycan-expressing cells (NG2 glia) represent a third subtype of macroglia in the brain. Originally described as oligodendrocyte precursor cells, they feature several characteristics not expected from mere progenitor cells, including synaptic connections with neurons. There is accumulating evidence that the properties of NG2 glia differ between different brain regions and developmental stages. To further analyze this proposed heterogeneity, we studied electrophysiological properties, transcript and protein expression, distribution and proliferative capacity of NG2 glia during postnatal development, focusing on the hippocampus and corpus callosum. All NG2 glia displayed a ‘complex’ current pattern consisting of voltage- and time-dependent in- and outward currents. In juvenile mice, Kir current densities and rectification index were highly variable and on average significantly lower than in adult animals. Single cell RT-PCR analyses of electrophysiologically characterized cells demonstrated that different glial genes were expressed at variable extent, independent of developmental stage and genetic background. In the hippocampus proper and the corpus callosum, the density of NG2 glia was highest at postnatal days (P)10–12, decreased by ~50 % at P25–35 and then remained stable in adults (P80–90). Interestingly, co-expression of NG2 and S100β, a marker for mature astrocytes, increased from 7 % at P10–12 to 27 % at P25–35 in the hippocampus proper, and then dropped again in the stratum radiatum at P80–90. In the dentate gyrus and corpus callosum, co-expression of NG2 and S100β was very low (3 %) and constant throughout development. Age-related differences were also observed with Ki-67, a proliferation marker. In NG2 glia of the CA1 region, its expression decreased from 16 % at P10–12 to 9 % (P25–35) and then 3 % (P80–90). Triple-stainings revealed that Ki-67 was also expressed in 2–3 % of NG2/S100β-positive cells in the juvenile and mature stratum radiatum, indicating that the latter, in contrast to S100β-positive astrocytes, still host proliferative potential. Taken together, we found significant differences in transcript and protein expression, electrophysiological properties and proliferative capacity of NG2 glia in the mouse forebrain, suggesting the co-existence of several subpopulations of NG2 glia. Our data thus support the idea of a substantial regional and developmental heterogeneity in this subtype of macroglia.
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Acknowledgments
We wish to thank Simone Durry for expert technical assistance. Supported by the Deutsche Forschungsgemeinschaft (DFG) (Ro 2327/8-1; SE 774/6; STE 552/4).
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B. Moshrefi-Ravasdjani, P. Dublin and G. Seifert equally contributed to this study.
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Supplementary Fig. 1 a: Co-expression based cell grouping in juvenile (P9 – 12) NG2-EYFP (n = 23 cells) and C57BL6J (n = 21) mice. b: Similar analysis was performed with cells from adult mice (P40 – 60) (NG2-EYFP mice, n = 24 cells; C57BL6J mice, n = 21) (PDF 415 kb)
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Moshrefi-Ravasdjani, B., Dublin, P., Seifert, G. et al. Changes in the proliferative capacity of NG2 cell subpopulations during postnatal development of the mouse hippocampus. Brain Struct Funct 222, 831–847 (2017). https://doi.org/10.1007/s00429-016-1249-2
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DOI: https://doi.org/10.1007/s00429-016-1249-2